Cell wall teichoic acid as a reserve phosphate source in Bacillus subtilis.

1. Quantitative determination of the anionic polymers present in the walls of Bacillus subtilis var. niger organisms undergoing transition, in a chemostat culture, from either Mg2+-limitation to PO43−-limitation or K+-limitation to PO43−-limitation showed that teichuronic acid synthesis started immediately the culture became PO43−-limited and proceeded at a rate substantially faster than the rate of biomass synthesis. 2. Simultaneously, the cell-wall teichoic acid content diminished at a rate greater than that due to dilution by newly synthesized wall material, and fragments of teichoic acid and mucopeptide accumulated in the culture extracellular fluid. 3. Equally rapid reverse changes occurred when a PO43−-limited B. subtilis var. niger culture was returned to being Mg2+-limited. 4. It is concluded that in this organism both teichoic acid and teichuronic acid syntheses are expressions of a single genotype, and a mechanism for the control of synthesis of both polymers is suggested. 5. These results are discussed with reference to the constantly changing environmental conditions that obtain in a batch culture and the variation in bacterial cell-wall composition that is reported to occur throughout the growth cycle.

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Genome-Wide Analysis of Phosphorylated PhoP Binding to Chromosomal DNA Reveals Several Novel Features of the PhoPR-Mediated Phosphate Limitation Response in Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.02570-14 ◽

2015 ◽

Vol 197 (8) ◽

pp. 1492-1506 ◽

Cited By ~ 14

Author(s):

Letal I. Salzberg ◽

Eric Botella ◽

Karsten Hokamp ◽

Haike Antelmann ◽

Sandra Maaß ◽

...

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Response Regulator ◽

Teichoic Acid ◽

Phosphate Limitation ◽

Chromosomal Dna ◽

Cell Wall Metabolism ◽

Wall Teichoic Acid ◽

Content Type ◽

Two Component

ABSTRACTThe PhoPR two-component signal transduction system controls one of three responses activated byBacillus subtilisto adapt to phosphate-limiting conditions (PHO response). The response involves the production of enzymes and transporters that scavenge for phosphate in the environment and assimilate it into the cell. However, inB. subtilisand some otherFirmicutesbacteria, cell wall metabolism is also part of the PHO response due to the high phosphate content of the teichoic acids attached either to peptidoglycan (wall teichoic acid) or to the cytoplasmic membrane (lipoteichoic acid). Prompted by our observation that the phosphorylated WalR (WalR∼P) response regulator binds to more chromosomal loci than are revealed by transcriptome analysis, we established the PhoP∼P bindome in phosphate-limited cells. Here, we show that PhoP∼P binds to the chromosome at 25 loci: 12 are within the promoters of previously identified PhoPR regulon genes, while 13 are newly identified. We extend the role of PhoPR in cell wall metabolism showing that PhoP∼P binds to the promoters of four cell wall-associated operons (ggaAB,yqgS,wapA, anddacA), although none show PhoPR-dependent expression under the conditions of this study. We also show that positive autoregulation ofphoPRexpression and full induction of the PHO response upon phosphate limitation require PhoP∼P binding to the 3′ end of thephoPRoperon.IMPORTANCEThe PhoPR two-component system controls one of three responses mounted byB. subtilisto adapt to phosphate limitation (PHO response). Here, establishment of the phosphorylated PhoP (PhoP∼P) bindome enhances our understanding of the PHO response in two important ways. First, PhoPR plays a more extensive role in adaptation to phosphate-limiting conditions than was deduced from transcriptome analyses. Among 13 newly identified binding sites, 4 are cell wall associated (ggaAB,yqgS,wapA, anddacA), revealing that PhoPR has an extended involvement in cell wall metabolism. Second, amplification of the PHO response must occur by a novel mechanism since positive autoregulation ofphoPRexpression requires PhoP∼P binding to the 3′ end of the operon.

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Analysis of Bacillus subtilis tagAB andtagDEF Expression during Phosphate Starvation Identifies a Repressor Role for PhoP∼P

Journal of Bacteriology ◽

10.1128/jb.180.3.753-758.1998 ◽

1998 ◽

Vol 180 (3) ◽

pp. 753-758 ◽

Cited By ~ 58

Author(s):

Wei Liu ◽

Stephen Eder ◽

F. Marion Hulett

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Phosphate Starvation ◽

Acid Synthesis ◽

Pho Regulon ◽

Wall Teichoic Acid ◽

Negative Role ◽

First Time ◽

Starvation Conditions

ABSTRACT The tagAB and tagDEF operons, which are adjacent and divergently transcribed, encode genes responsible for cell wall teichoic acid synthesis in Bacillus subtilis. TheBacillus data presented here suggest that PhoP and PhoR are required for direct repression of transcription of the two operons under phosphate starvation conditions but have no regulatory role under phosphate-replete conditions. These data identify for the first time that PhoP∼P has a negative role in Pho regulon gene regulation.

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Wall Teichoic Acid Polymers Are Dispensable for Cell Viability in Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.01336-06 ◽

2006 ◽

Vol 188 (23) ◽

pp. 8313-8316 ◽

Cited By ~ 123

Author(s):

Michael A. D'Elia ◽

Kathryn E. Millar ◽

Terry J. Beveridge ◽

Eric D. Brown

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Bacillus Subtilis ◽

Cell Viability ◽

Teichoic Acid ◽

Acid Synthesis ◽

Extensive Literature ◽

Wall Teichoic Acid ◽

First Time ◽

Similar Gene

ABSTRACT An extensive literature has established that the synthesis of wall teichoic acid in Bacillus subtilis is essential for cell viability. Paradoxically, we have recently shown that wall teichoic acid biogenesis is dispensable in Staphylococcus aureus (M. A. D'Elia, M. P. Pereira, Y. S. Chung, W. Zhao, A. Chau, T. J. Kenney, M. C. Sulavik, T. A. Black, and E. D. Brown, J. Bacteriol. 188:4183-4189, 2006). A complex pattern of teichoic acid gene dispensability was seen in S. aureus where the first gene (tarO) was dispensable and later acting genes showed an indispensable phenotype. Here we show, for the first time, that wall teichoic acid synthesis is also dispensable in B. subtilis and that a similar gene dispensability pattern is seen where later acting enzymes display an essential phenotype, while the gene tagO, whose product catalyzes the first step in the pathway, could be deleted to yield viable mutants devoid of teichoic acid in the cell wall.

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Phage SPP1 Reversible Adsorption to Bacillus subtilis Cell Wall Teichoic Acids Accelerates Virus Recognition of Membrane Receptor YueB

Journal of Bacteriology ◽

10.1128/jb.00349-08 ◽

2008 ◽

Vol 190 (14) ◽

pp. 4989-4996 ◽

Cited By ~ 84

Author(s):

Catarina Baptista ◽

Mário A. Santos ◽

Carlos São-José

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Strong Dependence ◽

Teichoic Acid ◽

Wall Teichoic Acid ◽

Binding Interaction ◽

Host Cell Membrane ◽

Teichoic Acids ◽

Bacterial Surface ◽

Reversible Adsorption

ABSTRACT Bacteriophage SPP1 targets the host cell membrane protein YueB to irreversibly adsorb and infect Bacillus subtilis. Interestingly, SPP1 still binds to the surface of yueB mutants, although in a completely reversible way. We evaluated here the relevance of a reversible step in SPP1 adsorption and identified the receptor(s) involved. We show that reversible adsorption is impaired in B. subtilis mutants defective in the glucosylation pathway of teichoic acids or displaying a modified chemical composition of these polymers. The results indicate that glucosylated poly(glycerolphosphate) cell wall teichoic acid is the major target for SPP1 reversible binding. Interaction with this polymer is characterized by a fast adsorption rate showing low-temperature dependence, followed by a rapid establishment of an equilibrium state between adsorbed and free phages. This equilibrium is basically determined by the rate of phage dissociation, which exhibits a strong dependence on temperature compatible with an Arrhenius law. This allowed us to determine an activation energy of 22.6 kcal/mol for phage release. Finally, we show that SPP1 reversible interaction strongly accelerates irreversible binding to YueB. Our results support a model in which fast SPP1 adsorption to and desorption from teichoic acids allows SPP1 to scan the bacterial surface for rapid YueB recognition.

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GtcA is required for LTA glycosylation in Listeria monocytogenes serovar 1/2a and Bacillus subtilis

10.1101/2019.12.12.873851 ◽

2019 ◽

Cited By ~ 2

Author(s):

Jeanine Rismondo ◽

Talal F. M. Haddad ◽

Yang Shen ◽

Martin J. Loessner ◽

Angelika Gründling

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Listeria Monocytogenes ◽

Teichoic Acid ◽

Lipoteichoic Acid ◽

Wall Teichoic Acid ◽

Gram Positive Bacteria ◽

Cell Wall Polymers ◽

Galactose Residue

ABSTRACTThe cell wall polymers wall teichoic acid (WTA) and lipoteichoic acid (LTA) are often modified with glycosyl and D-alanine residues. Recent studies have shown that a three-component glycosylation system is used for the modification of LTA in several Gram-positive bacteria including Bacillus subtilis and Listeria monocytogenes. In the L. monocytogenes 1/2a strain 10403S, the cytoplasmic glycosyltransferase GtlA is thought to use UDP-galactose to produce the C55-P-galactose lipid intermediate, which is transported across the membrane by an unknown flippase. Next, the galactose residue is transferred onto the LTA backbone on the outside of the cell by the glycosyltransferase GtlB. Here we show that GtcA is necessary for the glycosylation of LTA in L. monocytogenes 10403S and B. subtilis 168 and we hypothesize that these proteins act as C55-P-sugar flippases. With this we revealed that GtcA is involved in the glycosylation of both teichoic acid polymers in L. monocytogenes 10403S, namely WTA with N-acetylglucosamine and LTA with galactose residues. These findings indicate that the L. monocytogenes GtcA protein can act on different C55-P-sugar intermediates. Further characterization of GtcA in L. monocytogenes led to the identification of residues essential for its overall function as well as residues, which predominately impact WTA or LTA glycosylation.GRAPHICAL ABSTRACT

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Analysis of teichoic acid biosynthesis regulation reveals that the extracytoplasmic function sigma factor σ M is induced by phosphate depletion in Bacillus subtilis W23

Microbiology ◽

10.1099/mic.0.28021-0 ◽

2005 ◽

Vol 151 (9) ◽

pp. 3041-3049 ◽

Cited By ~ 11

Author(s):

Kathrin Minnig ◽

Vladimir Lazarevic ◽

Blazenka Soldo ◽

Catherine Mauël

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Regulatory System ◽

Phosphate Starvation ◽

Phosphate Limitation ◽

Wall Structure ◽

Wall Teichoic Acid ◽

Activity Data ◽

Two Component

The expression of the Bacillus subtilis W23 tar genes specifying the biosynthesis of the major wall teichoic acid, the poly(ribitol phosphate), was studied under phosphate limitation using lacZ reporter fusions. Three different regulation patterns can be deduced from these β-galactosidase activity data: (i) tarD and tarL gene expression is downregulated under phosphate starvation; (ii) tarA and, to a minor extent, tarB expression after an initial decrease unexpectedly increases; and (iii) tarO is not influenced by phosphate concentration. To dissect the tarA regulatory pattern, its two promoters were analysed under phosphate limitation: The P tarA -ext promoter is repressed under phosphate starvation by the PhoPR two-component system, whereas, under the same conditions, the P tarA -int promoter is upregulated by the action of an extracytoplasmic function (ECF) σ factor, σ M. In contrast to strain 168, σ M is activated in strain W23 in phosphate-depleted conditions, a phenomenon indirectly dependent on PhoPR, the two-component regulatory system responsible for the adaptation to phosphate starvation. These results provide further evidence for the role of σ M in cell-wall stress response, and suggest that impairment of cell-wall structure is the signal activating this ECF σ factor.

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Wall turnover deficiency of Bacillus subtilis Nil5 is due to a decrease in teichoic acid

Canadian Journal of Microbiology ◽

10.1139/m87-096 ◽

1987 ◽

Vol 33 (6) ◽

pp. 566-568 ◽

Cited By ~ 2

Author(s):

Ljubiša Vitković

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Stationary Phase ◽

Phosphorus Content ◽

Teichoic Acid ◽

Exponential Phase ◽

Standard Strain ◽

Late Exponential Phase ◽

Wall Teichoic Acid

Bacillus subtilis Ni15 is deficient in cell wall turnover. This deficiency is removed if the medium contains 0.2 M NaCl, which does not affect growth. The levels of amidase and glucosaminidase, the most likely enzymes involved in turnover, were, in stationary phase Nil5 cells, similar to those in late-exponential phase cells of a standard strain. The Nil5 enzymes were not salt sensitive. However, the Nil5 walls contained 4.7-fold less phosphorus than the walls of the standard strain. Since the phosphorus content of B. subtilis walls reflects the level of teichoic acid, it is proposed that the turnover deficiency of this strain is due to a decrease in wall teichoic acid.

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Teichoic Acid Is an Essential Polymer in Bacillus subtilis That Is Functionally Distinct from Teichuronic Acid

Journal of Bacteriology ◽

10.1128/jb.186.23.7865-7873.2004 ◽

2004 ◽

Vol 186 (23) ◽

pp. 7865-7873 ◽

Cited By ~ 68

Author(s):

Amit P. Bhavsar ◽

Laura K. Erdman ◽

Jeffrey W. Schertzer ◽

Eric D. Brown

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Wall Thickening ◽

Glycerol Phosphate ◽

Wall Teichoic Acid ◽

Teichoic Acids ◽

Gram Positive Bacteria ◽

Teichuronic Acid ◽

Wall Teichoic Acids

ABSTRACT Wall teichoic acids are anionic, phosphate-rich polymers linked to the peptidoglycan of gram-positive bacteria. In Bacillus subtilis, the predominant wall teichoic acid types are poly(glycerol phosphate) in strain 168 and poly(ribitol phosphate) in strain W23, and they are synthesized by the tag and tar gene products, respectively. Growing evidence suggests that wall teichoic acids are essential in B. subtilis; however, it is widely believed that teichoic acids are dispensable under phosphate-limiting conditions. In the work reported here, we carefully studied the dispensability of teichoic acid under phosphate-limiting conditions by constructing three new mutants. These strains, having precise deletions in tagB, tagF, and tarD, were dependent on xylose-inducible complementation from a distal locus (amyE) for growth. The tarD deletion interrupted poly(ribitol phosphate) synthesis in B. subtilis and represents a unique deletion of a tar gene. When teichoic acid biosynthetic proteins were depleted, the mutants showed a coccoid morphology and cell wall thickening. The new wall teichoic acid biogenesis mutants generated in this work and a previously reported tagD mutant were not viable under phosphate-limiting conditions in the absence of complementation. Cell wall analysis of B. subtilis grown under phosphate-limited conditions showed that teichoic acid contributed approximately one-third of the wall anionic content. These data suggest that wall teichoic acid has an essential function in B. subtilis that cannot be replaced by teichuronic acid.

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Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.00507-16 ◽

2016 ◽

Vol 198 (21) ◽

pp. 2925-2935 ◽

Cited By ~ 33

Author(s):

Heng Zhao ◽

Yingjie Sun ◽

Jason M. Peters ◽

Carol A. Gross ◽

Ethan C. Garner ◽

...

Keyword(s):

Bacillus Subtilis ◽

Transcriptional Repression ◽

Cell Envelope ◽

Turgor Pressure ◽

Teichoic Acid ◽

Lethal Gene ◽

Genetic Redundancy ◽

Wall Teichoic Acid ◽

Content Type ◽

Lipid Ii

ABSTRACTThe integrity of the bacterial cell envelope is essential to sustain life by countering the high turgor pressure of the cell and providing a barrier against chemical insults. InBacillus subtilis, synthesis of both peptidoglycan and wall teichoic acids requires a common C55lipid carrier, undecaprenyl-pyrophosphate (UPP), to ferry precursors across the cytoplasmic membrane. The synthesis and recycling of UPP requires a phosphatase to generate the monophosphate form Und-P, which is the substrate for peptidoglycan and wall teichoic acid synthases. Using an optimizedclusteredregularlyinterspacedshortpalindromicrepeat (CRISPR) system with catalytically inactive (“dead”)CRISPR-associated protein9(dCas9)-based transcriptional repression system (CRISPR interference [CRISPRi]), we demonstrate thatB. subtilisrequires either of two UPP phosphatases, UppP or BcrC, for viability. We show that a third predicted lipid phosphatase (YodM), with homology to diacylglycerol pyrophosphatases, can also support growth when overexpressed. Depletion of UPP phosphatase activity leads to morphological defects consistent with a failure of cell envelope synthesis and strongly activates the σM-dependent cell envelope stress response, includingbcrC, which encodes one of the two UPP phosphatases. These results highlight the utility of an optimized CRISPRi system for the investigation of synthetic lethal gene pairs, clarify the nature of theB. subtilisUPP-Pase enzymes, and provide further evidence linking the σMregulon to cell envelope homeostasis pathways.IMPORTANCEThe emergence of antibiotic resistance among bacterial pathogens is of critical concern and motivates efforts to develop new therapeutics and increase the utility of those already in use. The lipid II cycle is one of the most frequently targeted processes for antibiotics and has been intensively studied. Despite these efforts, some steps have remained poorly defined, partly due to genetic redundancy. CRISPRi provides a powerful tool to investigate the functions of essential genes and sets of genes. Here, we used an optimized CRISPRi system to demonstrate functional redundancy of two UPP phosphatases that are required for the conversion of the initially synthesized UPP lipid carrier to Und-P, the substrate for the synthesis of the initial lipid-linked precursors in peptidoglycan and wall teichoic acid synthesis.

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